Powerplex esx 17

DNA was extracted from the LCL-U-DCS cell line of the patient and from the U-DCS cell line at different passages, using the magnetic bead-based Maxwell-DNA IQ extraction protocol (Promega, Mannheim Germany). DNA extracts were diluted in 50 μl extraction buffer. PCR amplification was carried out with the self-developed Q11-STR kit, and two commercial STR kits, MPX-4 (Serac, Bad Homburg, Germany) and PowerPlex ESX 17 (Promega, Mannheim, Germany) according to the manufacturer’s recommendations. The three STR kits Q11, MPX4, and PowerPlex ESX 17 included the same STR markers: D3S1358, FGA, D8S1179, D18S51, D21S11, TH01, VWA, SE33, D2S1338, D16S539, D19S433, and the gender-typing system Amelogenin. PowerPlex ESX 17 contained the additional STRs D10S1248, D1S1656, D22S1045, D2S441, and D12S391. These kits are commonly used in forensic investigations for human identification and provide a very high probability of identity of 1 in 10¹³ (Q11 and MPX4) and 1 in 10¹⁸ (PowerPlex ESX 17).
Electrophoresis of the PCR products was carried out on an ABI Prism 3130 Genetic Analyzer (Applied Biosystems, Darmstadt, Germany) with the polymer POP7. The data was analyzed using the GeneMapper ID Software v3.2.

PCR was performed using a PowerPlex ESX 17 and PowerPlex HS 16 kit (Promega Corporation, USA) in a Gene Amp PCR System 9700 thermocycler (Applied Biosystems, USA). Amplification products were separated towards DNA CC5 ILS 500 and CC5 ILS 600 standards (Promega Corporation, USA) using a 3130 Genetic Analyzer (Applied Biosystems, USA). The following loci were analyzed: AMEL, D3S1358, TH01, D21S11, D18S51, D10S1248, D1S1656, D2S1338, D16S539, D22S1045, VWA, D8SS1179, FGA, D2S441, D12S391, D19S433, SE33, D5S818, D13S317, D7S820, TPOX, CSF1PO, Penta D, and Penta E. Additionally, alleles from chromosome Y were determined using a Yfiler test (Applied Biosystems, USA). PCR products were analyzed in a 3130 Genetic Analyzer (Applied Biosystems, USA). Genotypes were generated using Gene Mapper ID v3.2 software (Applied Biosystems, USA). Multiplex PCR and capillary electrophoresis procedures were executed according to the manufacturer’s recommendations.

DNA was extracted using The AutoMate ExpressTM Forensic DNA Extraction System with the PrepFilerTM Express BTA kit (Thermo Fisher Scientific, Darmstadt, Germany) to a 50-μL elution volume according to manufacturer’s instructions. Fluorometric DNA quantitation was performed using the Quantus™ Fluorometer with the QuantiFluor® dsDNA System (Promega, Walldorf, Germany) according to manufacturer’s instructions.
Additionally, triplicates of a human clavicle were processed according to the pig bone samples and stored for 10, 20, and 30 days and 35 °C in the substrate shown to result in the highest DNA recovery in pig bone. To check for STR profile quality, DNA from the positive control as well as the stored samples was amplified using the PowerPlex® ESX 17 (Promega, Mannheim, Germany) and the GlobalFiler™ PCR kits (Thermo Fisher Scientific, Darmstadt, Germany) according to manufacturer’s recommendations. The samples were analyzed using the 3130 Genetic Analyzer with the GeneMapper® ID software by Thermo Fisher Scientific.